棉蚜对四种棉花品种的选择性及GhRAC6基因的克隆研究

发布时间：2018-02-14 21:02

本文关键词： 棉蚜选择性验非选择性试验可溶性糖 SOD含量胼胝质 GhRac6基因　出处：《长江大学》2016年硕士论文　论文类型：学位论文

【摘要】：棉花具有重要的经济价值,是主要的纺织工业原料之一,但棉花生产中易受病虫害的危害,严重影响其自身的产量和品质,造成重大的经济损失。棉蚜是危害棉花的世界性害虫之一,通过刺吸式口器吸食植物韧皮部的营养物质,在取食植物汁液的同时时还会通过唾液传播植物病害。对于害虫的防治目前主要依赖化学农药,但化学防治不仅不利于环保,而且随着使用次数的增加,害虫会对其产生抗性。因此,培育高效抗虫的优质棉花一直是育种学家追求的目标之一。故本课题以棉花-棉蚜为研究对象,首先对四个棉花品种的抗蚜性进行了研究。在此基础上,通过生物信息学分析,分离鉴定了棉花的GhRac6基因。研究结果如下：1.无论是在选择性验中还是非选择性试验中,在接虫24h、48h、72h后,棉蚜对红叶矮的取食倾向都是最弱的,因此,初步断定红叶矮具有较强的抗蚜性。2.本试验对照与处理的4种棉花可溶性糖含量变化幅度大小为：徐州142中植棉红叶矮常国红叶棉。当4种棉花在接棉蚜48小时后,其体内的可溶性糖含量变化幅度最大的是徐州142,最小的是常国红叶棉,其含量变化幅度与前面棉蚜选择性和非选择性试验结果并不完全一致,王洪亮在棉蚜在不同品种上的发育历期和种群数量动态试验中也发现,可溶性糖含量的高低并不影响蚜虫的生长。该结论也可用王兴亚的观点解释,即：蚜虫危害大豆叶片与对照大豆叶片的可溶性糖含量无显著差异是由于蚜虫侵害大豆叶片取食糖份,使植物增加的可溶性糖量与蚜虫取食量达到了平衡,因此变化幅度较小。3.本试验对照与处理的4种棉花SOD含量变化幅度大小为：中植棉红叶矮常国红叶棉徐州142。该结论与前面接棉蚜48小时后,选择性和非选择性试验结果并不完全一致,这是由于SOD酶的调控机制是复杂的,Cu/Zn-SOD、Mn-SOD基因的表达量与其酶活性变化趋势并不完全一致,说明还有其它基因参与酶活性的调控。4.接棉蚜48小时后对照与处理的4种棉花叶片胼胝质相对增加量的大小为：徐州142中植棉常国红叶棉红叶矮。徐州142与红叶矮的结论也与前面的棉蚜的选择性和非选择性试验的结论表现出一致性,常国红叶棉与中植棉的相对增加量与前面的棉蚜的选择性和非选择性试验的结论不一致性,是由于棉花的抗性机制是复杂的,单一的胼胝质指标并不能代表抗虫的强弱。5.克隆了GhRac6基因,该基因由636bp碱基组成,分子量为23338.97D,等电点为6.718,编码211个氨基酸,并对其氨基酸的序列进行了同源分析,棉花RAC蛋白的系统进化树表明棉花基因GhRac6与陆地棉ADY68837.1亲缘关系最近。同时构建了植物表达载体pMD-GhRac6载体,为后续研究GhRac6的功能奠定了良好基础。
[Abstract]:Cotton has important economic value and is one of the main raw materials in textile industry. However, cotton production is vulnerable to pests and diseases, which seriously affects its own output and quality. The cotton aphid is one of the worldwide pests that harm cotton, feeding the nutrients in the phloem of the plant through the sucking mouthpiece. Plant diseases are transmitted through saliva while feeding on plant sap. At present, the control of pests mainly depends on chemical pesticides, but chemical control is not only harmful to environmental protection, but also increases with the increase of use times. Therefore, it has always been one of the objectives of breeders to develop high quality cotton with high efficiency and resistance to insects. Therefore, the subject of this study is cotton aphids, aphids, aphids, aphids, aphids, aphids, and aphids. Firstly, the resistance of four cotton varieties to aphid was studied. On the basis of this, the GhRac6 gene of cotton was isolated and identified by bioinformatics analysis. The results are as follows: 1. Whether in selective test or non-selective test, The feeding tendency of cotton aphids to red leaf dwarf was the weakest after 24 hours and 48 hours, so, It was preliminarily concluded that red leaf dwarf had strong resistance to aphid. The change range of soluble sugar content of four cotton varieties in the control and treatment was as follows: cotton grown in Xuzhou 142, red leaf dwarf red leaf cotton, when four cotton species were inoculated with cotton aphid for 48 hours. The content of soluble sugar in Xuzhou 142was the largest, and the smallest in Changguo red leaf cotton. The change range of soluble sugar content was not completely consistent with the results of selective and non-selective experiments on aphid aphid. Wang Hongliang also found that the content of soluble sugar did not affect the growth of aphids in the development period and population dynamics of different varieties of cotton aphid. This conclusion can also be explained by Wang Xingya's viewpoint. That is to say, there was no significant difference in soluble sugar content between soybean leaves and control soybean leaves caused by aphids, because aphids invaded soybean leaves to eat sugar, so that the increased soluble sugar content of plants and the feeding amount of aphids reached a balance. Therefore, the variation range of SOD content in the four cotton varieties of the control and treatment was as follows: middle cotton with red leaves, red leaf Dwarf, red leaf cotton, Xuzhou, Xuzhou, 142.This conclusion was compared with that of aphid aphids in front of cotton aphid for 48 hours. The results of selective and non-selective tests are not completely consistent, which is due to the complex regulatory mechanism of SOD enzyme, which is not completely consistent with the changing trend of the expression of Cu- / Zn-SODn-SOD gene and its enzyme activity. It shows that there are other genes involved in the regulation of enzyme activity .4.After 48 hours of inoculation with APHIS gossypii, the relative increase of callosum in the leaves of four cotton varieties treated with APHIS gossypii is as follows: the relative increment of callosum of cotton leaves in Xuzhou 142. The short conclusion was also consistent with the results of the previous selective and non-selective tests of the cotton aphid (APHIS gossypii). The results of selective and non-selective tests of cotton aphids and cotton aphids were inconsistent because of the complex resistance mechanism of cotton. The GhRac6 gene was cloned from 636bp base group with a molecular weight of 23338.97D and a isoelectric point of 6.718, encoding 211 amino acids. The phylogenetic tree of cotton RAC protein indicates that the cotton gene GhRac6 is closely related to Upland cotton ADY68837.1. Meanwhile, a plant expression vector pMD-GhRac6 vector was constructed, which laid a good foundation for further study on the function of GhRac6.
【学位授予单位】：长江大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.622.1

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